Bowen-Conradi syndrome (BCS) is an autosomal-recessive disorder characterized by severely impaired prenatal and postnatal growth, profound psychomotor retardation, and death in early childhood. Nearly all reported BCS cases have been among Hutterites, with an estimated birth prevalence of 1/355. We previously localized the BCS gene to a 1.9 Mbp interval on human chromosome 12p13.3. The 59 genes in this interval were ranked as candidates for BCS, and 35 of these, including all of the best candidates, were sequenced. We identified variant NM_006331.6:c.400A-->G, p.D86G in the 18S ribosome assembly protein EMG1 as the probable cause of BCS. This mutation segregated with disease, was not found in 414 non-Hutterite alleles, and altered a highly conserved aspartic acid (D) residue. A structural model of human EMG1 suggested that the D86 residue formed a salt bridge with arginine 84 that would be disrupted by the glycine (G) substitution. EMG1 mRNA was detected in all human adult and fetal tissues tested. In BCS patient fibroblasts, EMG1 mRNA levels did not differ from those of normal cells, but EMG1 protein was dramatically reduced in comparison to that of normal controls. In mammalian cells, overexpression of EMG1 harboring the D86G mutation decreased the level of soluble EMG1 protein, and in yeast two-hybrid analysis, the D86G substitution increased interaction between EMG1 subunits. These findings suggested that the D-to-G mutation caused aggregation of EMG1, thereby reducing the level of the protein and causing BCS.

Cyprinid herpesvirus 3 (CyHV-3) is the aetiological agent of koi herpesvirus disease in koi and common carp. The disease is notifiable to the World Organisation for Animal Health. Three tests -quantitative polymerase chain reaction (qPCR), conventional PCR (cPCR) and virus isolation by cell culture (VI) -were validated to assess their fitness as diagnostic tools for detection of CyHV-3. Test performance metrics of diagnostic accuracy were sensitivity (DSe) and specificity (DSp). Repeatability and reproducibility were measured to assess diagnostic precision. Estimates of test accuracy, in the absence of a gold standard reference test, were generated using latent class models. Test samples originated from wild common carp naturally exposed to CyHV-3 or domesticated koi either virus free or experimentally infected with the virus. Three laboratories in Canada participated in the precision study. Moderate to high repeatability (81 to 99%) and reproducibility (72 to 97%) were observed for the qPCR and cPCR tests. The lack of agreement observed between some of the PCR test pair results was attributed to cross-contamination of samples with CyHV-3 nucleic acid. Accuracy estimates for the PCR tests were 99% for DSe and 93% for DSp. Poor precision was observed for the VI test (4 to 95%). Accuracy estimates for VI/qPCR were 90% for DSe and 88% for DSp. Collectively, the results show that the CyHV-3 qPCR test is a suitable tool for surveillance, presumptive diagnosis and certification of individuals or populations as CyHV-3 free.KEY WORDS: CyHV-3 · Diagnostic validation · Precision · Accuracy · Quantitative PCR · Virus isolation Resale or republication not permitted without written consent of the publisherDis Aquat Org 123: 2017 ment on the Application of Sanitary and Phytosanitary Measures.An effective CyHV-3 diagnostic assay should detect the virus in diseased fish as well as in asymptomatic carriers of the virus with latent or persistent infections. Three diagnostic assays were evaluated for their performance characteristics in this study: a modified conventional polymerase chain reaction (cPCR) assay targeting the CyHV-3 thymidine kinase gene (cTK;Bercovier et al. 2005), a modified quantitative PCR (qPCR) assay targeting CyHV-3 open reading frame 89 (ORF89) (qORF89; Gilad et al. 2004) and virus isolation by culture on common carp brain (CCB) cells (VI;Neukirch et al. 1999). The unmodified cTK and qORF89 tests have been evaluated in inter-laboratory ring studies (Way 2008) and intralaboratory comparisons (Bergmann et al. 2010, Monaghan et al. 2015a). The qPCR test displayed high analytical specificity (ASp), detecting all known CyHV-3 isolates but not related virus species such as CyHV-1 or CyHV-2 (Gilad et al. 2004, Bergmann et al. 2010. It is currently the most commonly used diag nostic test for CyHV-3 and may prove to be suitable for use in surveillance programs to declare healthy populations of susceptible fish free of the virus (OIE 2015c). The cTK test is one of the PCR assays recommended by the OIE fo...

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Four new ruthenium(II) complexes [Ru(mpt)2(DMSO)2] (1), [Ru(mpt)2(bpy)] (2), [Ru(mpt)2(phen)] (3) and [Ru(mpt)2(tptz)] (4) have been synthesized and characterized by elemental analyses, IR, 1H and 13C NMR, and electronic absorption spectroscopy....